6alpha-cyano progesterone and derivatives thereof



Patented Jan. 30, 1962 United States Patent Ofitice 3701924 3,019,240 Goc-CYANO PROGESTERONE AND DERIVATIVES THEREOF Albert Bowers and Howard J. Ringold, Mexico City, Mexico, assignors to'Syntex, S.A., Mexico City, Mexico, a corporation of Mexico N Drawing. Filed Nov. 5, 1959, Ser. No. 850,595

application Mexico Nov. 6, 1958 Claims priority,

27 Claims. (til. 260-3973) in which Y is selected from the group consisting of.

0 H and X is a member of the group consisting of hydrogen and fluorine and is hydro-gen where Y is hydrogen, and fluorine where Y is =0 about 12 carbon atoms; the hydrocarbon carboxylic acid can be saturated or unsaturated, of straight, branched, cyclic or mixed aliphatic-cyclic chain, which may have functional substitucnts such as hydroxyl, acyloxy (of 1 to 12 C), all oxy (1 to 5 C) or halogen; typical esters derived from such acids are the acetate, propi onate', isobutyrate, hemisuccinate, enanthate, caproate, benzoate, trimethylacetate, phenoxyacetate, phenylpropionate and fl-chloropropionate. I

Our invention also comprises the novel 6-cyano derivatives of l9-nor-progesterone and l7a-hydroxy-19-norprogesterone, represented by the general formulas:

CHa

| l C O wherein R, X and Y have the same meaning asfset forth above.

The new compounds beingthe object of the present invention and particularly those which have no substituent at Cell are potent proge's tation-al agents of anti-androgenic-and anti-estrogenic activity; the ll-substitutedcompounds of the IO-methyl series are in addition useful intermediates in the synthesis of 6-cyano cortical hormones which are anti-inflammatory agents and may readily be prepared from the former compounds by conventional in troduction of the 2l-hydroxy group."

The process for making the new compounds according to the invention essentially involves the introduction of a I 6a-cyano group into the starting compounds and, if del sired, the'subsequent introduction bonds mentioned above.

of the additional double I The aforesaid process can be illustrated by the following reaction diagram, in which R, X and; Y have the same meaning as explained hereinbefore, while E is a member of the group consisting of methyl and hydrogen;

dehydrogenation dehydrogenation The starting compounds (I) were first converted to their 3,20-bis-ethylenedioxy derivatives (IA) and then to their 3,ZO-bis-ethylenedioxy-Sabet-oxide derivatives (1B) and these latter derivatives were refluxed with potassium cyanide in mixture with ethyleneglycol. This reaction afforded a mixture of the respective 6-cyano-3,20 bisethylenedioxy-A -pregnene and of the corresponding 6- cyano 3 hydroxyethoxy-ZO-ethylenedioxy-A -pregnadicue; the mixture was directly treated with dry hydrogen chloride in glacial acetic acid solution thereby produc- 10 ing the 6a-cyano-compounds (II) of the lO-methyl series,

or those of the 19-nor series (IIA) depending on the starting material used, in equilibrium with their 3-enol form.

Optionally, we then introduced into the compounds (II) a double bond at C-1 by refluxing with selenium dioxide in mixture with t-butanol and in the presence of catalytic amounts of pyridine (III).

We introduced a double bond at C-6 into either compound II or III, for which purpose a solution of the 20 steroid in t-butanol was, for instance, treated with approximately 1.1 molar equivalents of sodium metlioxide, and the resulting 6-eyano-6-sodio compound, without isolation, was treated with 1 molar equivalent of bromine to produce a 6-bromo-6-cyano compound which was then dehydrobrorninated by refluxing with -c0llidine or with a mixture of calcium carbonate and dimethylformamide. Thus we produced the 6-cyano-6-dehydro compounds (IV) or the 6-cyano 1,6-bis-dehydro compounds (V).

By a similar method to that described above we also introduced the double bond at C-6 into 6a-cyan0-19-norldehydrogenntion to AA) OHa (His

(1311: CO ---R CHg progesterona and 60a cyano-l9-nor-17a-hydroxypro gesterone (IIA) as well as. into their ll-oxygenated derivatives' to obtain compounds (IVA).

For esterifying thehydroxyl group at C-17 we employed conventional methods of esterification. In the case of the acetylations with acetic anhydride and acetyl chloride we also isolated 3,17-diacetates of the corresponding intermediate enol.

The reactions described above and more in detail in the examples given hereinafter may be modified within wide limits, both with respect to the reagents and solvents employed as with respect to the conditions of temperature and time. To name some of the possible modifications: the dehydrogenation with selenium dioxide may i also be effected in another solvent such as acetic acid, in the presence or in the absence of an adequate catalyst, such as water; instead of t-butanol there may be used another tertiary aliphatic alcohol and/or the pyridine may be substituted by another basic catalyst such as -collidine; the refluxing period with selenium dioxide in t-butanol may fluctuate within wide limits, for example between 12 and 96 hours; the dehydrogenation at Example 1' In a conventional manner, for instance by the method described by S. Bernstein et al. in J. Org. Chem. 17, p. 1341 (1952) we prepared the 3,20-bis-cycloethyleneketal of progesterone.

The epoxidation of the nuclear double bond of this his ketal was carried out as follows: a chloroform solution of the ketal (20 cc. of chloroform for 1 g. of steroid) was cooled to O C. and then treated with an ether solution of monoperphthalic acid containing 1.2 molar equivalents of the peracid; the mixture was kept at C. in the dark for 16 hours. organic layer was separated, washed with water, dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure. By chromatography of the residue on neutral alumina there was obtained 3,20

bis-cycloethylenedioxy-a,6a-oxido-pregnane.

A mixture of 5 g. 3,ZO-bis-ethylenedioxy-Sa,6x-oxidopregnane (J. Chem. Soc., 4112 (1957)), g. of potassium cyanide and 200 cc. of ethyleneglycol was refluxed for half an hour, poured into ice water and the precipitate formed was collected by filtration, washed with water and dried. The product consisted of a mixture of 6-cyano- 3,20-bis-ethylenedioxy-M-pregnene (M.P. 182-184 C.,

[ab-74, x 244mm, e=9,590) and of the S-hydroxyethylether of 6-cyano-20-ethylenedioxy-A -pregnadien- 3-01 (M.P. 175 177 C., [ab-96, x 282284m,u, e=23,990); a small amount of this mixture was separated into its components by chromatography on neutral alumina.

A slow stream of dry hydrogen chloride was introduced into a solution of the above mixture in 200 cc. of glacial acetic acid, for a period of 3 hours and maintaining the temperature below 18 C.; after pouring into ice water the precipitate was collected, washed with water, dried,

After dilutingwith water the all) There was Example I] A mixture of 3 g. of 6a-cyano-progesterone obtained as described in the preceding example, 100 cc. of t-butanol, 1.2 g. of recently sublimed selenium dioxide and 0.2 cc. of pyridine was refluxed under an atmosphere of nitrogen for 48 hours and then filtered through celite. The solvent was evaporated under reduced pressure, the residue was dissolved in acetone, treated with decolorizing charcoal, refluxed for 2 hours, filtered and the acetone was evaporated. The resulting residue was purified by chromatography on neutral alumina, thus yielding 6a-cyano-l-dehydro-progesterone.

A solution of 2 g. of the latter compound in 60 cc. of t-butanol was added to 1.1 molar equivalents of sodium :methoxide prepared by dissolving the corresponding amount of sodium metal in absolute methanol and evaporating the methanol under anhydrous conditions. The mixture was stirred at room temperature for 30 minutes and a solution of 1.1 molar equivalents of bromine in 30 cc. of t-butanol was then slowly added under stirring and while maintaining the temperature below C. The mixture was kept for half an hour at room temperature and diluted with water, and the formed precipitate was collected by filtration, washed with water and dried under vacuum. Crude 6-cyano-6-brorno-ldehydro-progesterone was obtained. By recrystallization of a small amount of this product from acetone-hexane at low temperaturethere was obtained the pure 6-cyano- 6bromo-l-dehydro-progesterone.

The'above crude compound was dissolved in cc. of dimethylformamide; the solution was added to a suspension of 1.5 g. or" calcium carbonate in cc. of dimethylformamide heated to boiling and the mixture was refluxed for 15 minutes, concentrated to about 20 cc. under reduced pressure, cooled and poured into aqueous saturated sodium chloride solution, and the precipitate was collected; washed with water, dried and recrystallized from acetonehexane. There was thus: obtained 6- cyano-l ,6-bis-dehydro-progesterone.

Example III In accordance with the method of Example I there were prepared 2 g. of 6a'-cyano-progesterone which was then treated with sodium methoxide, bromine and calcium carbonate, as described in Example II. There was thus obtained 6-cyano-6-dehydro-progesterone.

The latter was subjected to a reaction with selenium dioxide, exactly as described in the preceding example, to obtain 6-cyano-1, 6-bis-dehydro-progesterone, identical with the compoundobtained in accordance with the method described in Example II.

Example IV A mixture of 3 g. of 6-bromo-6-cyano-progesterone, obtained as an intermediate in'the preceding example, and cc. of 'y-collidine was refluxed for 1 hour, cooled and diluted with ether, and the collidine hydrobromide was removed by filtration; the filtrate was consecutively washed with dilute hydrochloric acid, 5% aqueous sodium carbonate solution and water, and dried over anhydrous sodium sulfate, and the ether was evaporated; recrystallization of the residue from acetone-hexane yielded 6-cyano-6-dehydro-progesterone, identical with the compound obtained as an intermediate in the method of the preceding example.

Example V By following the procedures describedin the preceding Example I the new compounds listed below were produced from the cited starting materials:

terone. 11- keto 9afiuoro progest rone. l1fl-hydroxy-9a-fiuore-progesterone.

11 keto 9a fluoro l7a-hydroxy-progestcrone.

19-nor-17a-hydroxyprogesterone.

11-keto-19-nor-pro- Fried et al., 1.11.0.8. 7711p. 1068 (1955). o

Well known and conventional.

Bowers et a1, Tetra- Starting Material Source New Product a... l7a-hydroxy-proges- Well known and 6a-cyano-17a-hydroxterone. conventional. y-progesterone. b. ll-keto-progesterone. do fim-eyano-ll-ketdprogesterone. c. IIB-hydroxy-progesdo fia-eyano-llfl-hydroxy-progesterone. fia-eyano-ll-keto-Qafluoro-progesterone 6a-eyano-l1fi-hydr0xy-9afluoro-progesterone. fia-eyano -11-keto-9afiuoro-17a-hydroxyprogesterone. fia-eyano-lQ-nor-progesterone. 6a-cyano-19-nor-17ahydroxy-progesterone. Gweyano-ll-keto-l gesterone. hedron, Vol. 2, p. nor-progesterone.

163 (1958). j 11B-hydroxy-19-nor- Bowers et al., Tetrafia cyano-llfl-hydroxprogesterone. hedron, Vol. 2, p. y-19-nor-progcster- 165 (1958). one. k. 9a fluoro 11!: hy prepared from the 60: eyano-Qa -fluor droxy-lQ-nor-progesterone.

known lla-hydroxy-lQ-nor-progesterone Pederson et al., J. Am. Chem. Soc, 78, 1512 (1956) by the method described in U. S. Patent 2,852,511.

115 hydroxy l 9 nor-progesterone.

Example V1 Example I was repeated with 11/3,17a-dihydroxy-19- nor-progesterone as the starting material and there was obtained 6tx'cyano-l1 8,l7a-dihydroxy-19-nor-progesterone.

The above starting material was prepared from conventional 17tz-hydroxy-19-nor-progesterone by the following method:

A culture of Curvularia lunata (Syntex strain 192) was prepared by inoculating an aqueous medium containing 2% by weight of peptone and of corn syrup with a vegetative growth of the aforesaid microorganism in the same medium and incubating the culture under stirring at a temperature of 28 C. and under aeration for about 24 hours.

To each liter of this culture there were added 30 cc. of an ethanolic solution of 17a-hydroxy-19-nor-progesterone having a concentration of 10 milligrams of the steroid per cc. The mixture was stirred under aeration at 28 C. for 24 hours. then extracted repeatedly with methylene chloride, the extract was washed with water, dried on sodium sulfate, filtered and the filtration concentrated to a small volume under reduced pressure. A total of 3 g. of 17u-hydroxy- 19-nor-progesterone were incubated in this manner. The combined concentrated extracts were then adsorbed on a column charged with 60 g. of silica gel and 60 g. of celite, previously washed with methylene chloride. The charged column was then eluted with a mixture of methylene chloride and acetone in a volume ratio of 4:1; the eluted 11j8,17a-dihydroxy 19 nor-progesterone was recovered from the eluate by evaporation of the solvent and the resulting residue was recrystallized from methylene chloride/ acetone so as to obtain pure 11,9,17a-dihydroxy-19- nor progesterone (A -19-nor-pregnene-11 13,17oc-di01-3,20-

dione) Example VII solved in 20 cc. of acetic acid and treated at 15 to 18 The incubation product was C. with a solution of 240 mg. (1.2 mol equivalents) of chromium trioxide in 10 cc. of acetic acid, the reaction mixture was left standing at room temperature for one hour. Then it was poured into ice Water, and the resulting precipitate collected, washed with water until neutral, and recrystallized from methylene chloride/ ether, thus yielding 11-keto-17a-hydroxy-19-nor-progesterone.

Example VIII By following the procedure of Examples II to V1, or a similar known dehydrogenation treatment, for instance with chloranil, there wer obtained the new A A and A derivatives listed below from the new C and C saturated compounds prepared as cited below:

New A A and A -derivatives fia-cyano-fla-hydroxy-l -dehydro-progesterone. 6-cyano-17a-hydroxy-G-dehydro-progesterone. fi-eyano-17a-hydroxy-1,6-bis-dehydro-progester- Example IX A mixture of 1 g. of Got-cyano-17a-hydroxyprogesterone, obtained as described in Example Va, 25 cc. of acetic anhydride and 5 cc, of acetyl chloride was refluxed for 3 hours under an atmosphere of nitrogen and the solvents were removed by distillation under reduced pressure, avoiding overheating. The residue was recrystallized from methanol-chloroform containing a few drops of pyridine. There was thus obtained the diacetate of 6-cyano-n -pregnadiene-3,17a-diol-20-one (M.P. 202- 203 C., [ab-112, )t 262-264mp, e=l8,620.

500 mg. of the above diacetate was treated with 25 cc. of a 1% methanolic solution of potassium hydroxide and stirred under an atmosphere of nitrogen for 1 hour at 0 C.; the mixture was acidified with acetic acid, concentrated to about 10 cc. and poured into 60 cc. of cold aqueous saturated sodium chloride solution; the precipitate was collected, washed with a little cold water, dried and recrystallized from acetone-hexane. There was thus obtained the acetate of 6a-cyano-l7u-hydroxyprogesterone.

Example X By the following the method described in the preceding example there was acetylated the 17ahydroxyl group of 6-cyauo-6-dehydro-l7a-hydroxyprogesterone, via the diacetate of 6-cyano-A -pregnatriene-B,17a-dio1-20-one.

Example XI A mixture of 1 g. of 6ot-cyano-17a-hydroxy-19-norprogesterone obtained according to Example Vh, 1 g. of p-toluenesulfonic acid monohydrate, 50 cc. of acetic acid and 10 cc. of acetic anhydride was allowed to react for 3 hours at room temperature. The mixture was then diluted with water, extracted with ether and the extract was consecutively Washed with 5% aqueous sodium carbonate solution and with aqueous saturated sodium chloride solution, dried over anhydrous sodium sulfate and the ether was evaporated. Crystallization of the residue Example XII A mixture of 1 g. of 6-cyano-6-dehydro-17a-hydroxyprogesterone, obtained according to Example VlIIb, 50 cc. of benzene, 2 g. of benzoic anhydride and 100 mg. of o-toluenesulfonic acid was kept at room temperature for 48 hours and then diluted With water; the benzene layer was repeatedly washed with Water and the solvent was evaporated under reduced pressure. The residue was treated with 50 cc. of 1% methanolic potassium hydroxide solution and the product was isolated as described in Example IX, thus affording the benzoate of 6-cyano- 6-dehydro-17a-hydroxyprogesterone.

By dehydrogenation with selenium dioxide, in accordance with the procedure described in Example II, there was obtained 6-cyano-1,6-bis-dehydro-l7a-hydroxyprogesterone benzoate.

Example XIII A mixture of 1 g. of 6-cyano-l,6-bis-dehydro-17ahydroxyprogesterone, obtained according to Example VIIIc, 50 cc. of benzene, 3 g. of cyclopentylpropionic anhydride and 200 mg. of p-toluenesulfonic acid was allowed to react at room temperature for 72 hours. The product Was then isolated in accordance with the method described in the preceding Example, thus yielding 6-cyano- 1,6-bis-dehydro-17a-hydroxyprogesterone cyclopentylpropionate.

We claim:

1. A new compound corresponding to a general formula selected from the group consisting of CH CH3 O on, IWR on,

(EN ON CH; CH;

(IJH; $13.: $0 0'30 ---R ---R /\i] X X H x l H and 0: O:

I on H in which formulas R is a member of the group consisting of hydrogen, hydroxy and the acyloxy radical of a hydrocarbon carboxylic acid having up to about 12 carbon atoms; in which Y is selected from the group consisting of H OH =0 and H X is a member of the group consisting of hydrogen and fluorine and is hydrogen where Y is hydrogen, and fluorine Where on Yis =0 or H and Z is a member of the group consisting of C-C and C=C.

6a-cyano-progesterone. 6a-cyano-l7a-hydroxy-progesterone. 6a-cyano-1l-keto-progesterone. 6ot-cyano-llfl-hydroxyprogesterone. 6a-cyano-l9-nor-progesterone. 6rx-cyano 19-nor-17tx-hydroxy-progresterone. 6ot-cyano 19-nor-ll-keto'progesterone.

6a-cyano 11a-hydroxy-l9-nor-progesterone. 6a-cyano 9ot-fiuoro-1l-keto-progesterone. GOL-CYHHO 9a-fluoro-1l s-hydroxy-progesterone. 6a-cyano-l-dehydro-progesterone. 6-cyano-1,6-bis-dehydro-progesterone. 6-cyano-6-dehydro-progesterone. Got-cyano-17whydroxy-l-dehydro-progesterone. 6-cyano-17a-hydroxy-6-dehydro-progesterone. 6-cyano-17a-hydroxy-l,6-bis-dehydro-progesterone.

6a-cyano-11-keto-1-dehydro-progesterone. 6-cyano-11-keto-6-dehydroprogesterone. 20. 6-cyano-1l-keto-1,6-bis-dehydro-progesterone. 21. 6u-cyano-9wfiuoro-l lfl-hydroxy-l-dehydro-progesterone.

22. 6-cyano-9a-fluoro-11/3-hydroxy-6-dehydro progesterone.

23. 6-cyano 9a fluoro-llfi-hydroxy-1,6-bis-dehydroprogesterone.

24. 6-cyano-6-dehydro-19-nor-progesterone. 25. 6-cyano-6-dehydro-17u-hydroXy-19nor progesterone.

26. 6-cyano-6-dehydro-l1p,17u-dihydroxy-19-nor progesterone.

27. 6-cyano-6-dehydro-1l-keto-lh-hydroxy 19 norprogesterone.

References Cited in the file of this patent UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. $019,240 January '30 1962 Albert Bowers et al5 corrected below.

Column 9 lines 45 to 59,, the Fight-Juana] formula should appear as shown below instead of as in the patent:

Signed and sealed this 31st dy of July 1962. I

SEA L) Attest:

ERNEST W. SWIDER 1 DAVID L. LADD Attesting Officer Commissioner of Patents 

1. A NEW COMPOUND CORRESONDING TO A GENERAL FORMULA SELECTED FROM THE GROUP CONSISTING OF 